Overview - ISO/TS 24420:2023 (shotgun metagenomic data processing)
ISO/TS 24420:2023 provides general requirements and guidelines for the data processing, quality control, storage, sharing and interoperability of shotgun metagenomic sequence data generated by massively parallel DNA sequencing. The technical specification covers workflows for host‑derived microbiomes and environmental metagenomes, specifying expectations for preprocessing, sequence QC, assembly, annotation and metadata organization. Note: functional analysis is explicitly excluded.
Key topics and technical requirements
- Scope and workflow: Defines the end‑to‑end metagenomic workflow - sequencing → data processing (preprocessing, QC, assembly) → data analysis (annotation, abundance calculation) → archiving/metadata.
- Quality control metrics: Requires initial QC of raw reads following ISO 20397 standards. Example numeric thresholds given in the specification: Q20 ≥ 90% and Q30 ≥ 80% for base quality metrics.
- Software and facilities:
- Bioinformatics pipelines must be validated and “locked down” (tools, code, runtime environment and network configuration). Any component changes require revalidation.
- High‑performance computing is recommended for large datasets and complex genome collections.
- Sequence assembly and annotation: Specifies processing steps for contigs/scaffolds and guidelines for annotation and calculation of species relative abundance (gene‑ and species‑level analyses).
- Data archive, directory and metadata:
- Guidance for organizing original/raw data, sequencing analytical data, and a structured directory of data elements for traceability and findability.
- Metadata requirements to support storage, sharing and interoperability; annexes provide examples of data formats and directory structures.
- Normative references: Ties into ISO 20397 series for nucleic acid and library preparation and QC procedures.
Practical applications and users
ISO/TS 24420:2023 is intended for:
- Clinical and research sequencing facilities performing shotgun metagenomic analyses.
- Bioinformatics teams and data managers implementing validated pipelines and quality assurance.
- Environmental microbiology and microbiome researchers standardizing data formats and metadata for sharing and reproducibility.
- Biotechnology companies, public health labs and data repositories aiming for interoperable metagenomic archives.
Practical benefits include improved data quality, reproducibility of metagenomic analyses, streamlined data sharing, and clearer requirements for pipeline validation and metadata curation.
Related standards
Keywords: ISO/TS 24420:2023, shotgun metagenomic, massively parallel DNA sequencing, quality control, metadata, data processing, sequence assembly, data archiving.
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